Application And Basic Research Of Stereotactic Technology With Frame In Parkinson’s Disease

Background and objective: Parkinson’s disease(PD)is a common neurodegenerative disease in middle-aged and elderly people.It not only brings physical and psychological pain to patients and families,but also brings heavy economic burden to society.Its pathological features are progressive loss of dopaminergic neurons in substantia nigra,decrease of dopamine content in striatum,aggregation of α-syn and over expression of neuroinflammation play an important role in Parkinson’s disease.Deep brain stimulation of subthalamic nucleus(STN-DBS)based on stereotactic technique with frame can significantly improve the motor symptoms of Parkinson’s disease,but the improvement of non motor symptoms is uncertain.How to improve the motor symptoms of patients and improve the non motor symptoms is the focus of many medical science and technology workers.In recent years,more and more attention has been paid to the role of microglial inflammation in the pathogenesis and regulation of Parkinson’s disease.Studies have shown that mi RNA plays a regulatory role in microglial neuritis induced by α-syn.Mir-let-7a is a very important member of the let-7 family and has been found to regulate microglial inflammation in vitro.We hypothesized that mir-let-7a may play a role in the pathogenesis of PD and try to explore its mechanism.We retrospectively analyzed 15 patients with advanced Parkinson’s disease who underwent STN-DBS surgery in Jiangxi Provincial People’s Hospital,in order to provide reference for future clinical treatment and improve the clinical operation effect of deep brain stimulation based on frame stereotactic technology.Meanwhile,we transplanted mir-let-7a mimics into a-syn overexpressed mouse model by stereotactic technique,objective to investigate the inhibitory effect of mir-let-7a on microglial inflammation induced by a-syn by targeting STAT3,hoping to explain the positive role of mir-let-7a in mouse Parkinson’s disease model.The study of this molecular mechanism may provide new targets and treatment strategies for the potential treatment of Parkinson’s disease,and provide an idea for the treatment of non motor symptoms of Parkinson’s disease.The first part: Study on deep brain stimulation of subthalamic nucleus based on framed stereotactic technique in Parkinson’s diseaseObjective: Deep brain stimulation(DBS)is a kind of stereotactic and precise operation.It can locate the target point through imaging method(CT or MRI),implant microelectrode and give electrical stimulation,so as to improve the motor symptoms and reduce the dose of levodopa.Deep brain stimulation of subthalamic nucleus can improve the motor symptoms such as static tremor,myotonia,motor retardation and dyskinesia.In view of its adjustable,safe and effective characteristics,deep brain stimulation is a better choice for the treatment of Parkinson’s disease in the middle and late stage.The non motor symptoms of Parkinson’s disease include fatigue,depression,anxiety,sleep disorder,constipation,bladder and other autonomic nervous dysfunction,sensory disorders,etc.the improvement effect of motor symptoms after the subthalamic nucleus deep brain stimulation(STN-DBS)is clear,and the improvement of non motor symptoms is uncertain.In this study,15 patients with Parkinson’s disease in the middle and late stage who were operated by STN-DBS in Jiangxi Provincial People’s hospital were analyzed retrospectively to evaluate the therapeutic effect of surgery,so as to provide reference for future clinical treatment and improve the therapeutic effect of deep brain electrical stimulation.Methods: Data of PD patients treated with bilateral STN-DBS from January 2017 to January 2019 in neurosurgery department of Jiangxi people’s hospital were collected.The deviation between the electrode tip and the planned target on X and Y axis was compared.The total score of UPDRS III of on-off-drug states in the early morning,the equivalent dose of L-dopa per day,the onset time of L-dopa shock test and the effective duration of L-dopa were collected before operation.The total scores of UPDRS III of sim on+drug off states,UPDRS III of stim on+drug on states,levodopa equivalent dose per day,the onset time of levodopa impact test and the duration of levodopa were collected one year after operation.The improvement of postoperative motor symptoms,the change of preoperative and postoperative cognitive and mental test scores,the equivalent dose of L-dopa per day and the effective time of L-dopa were compared.Results: 15 cases were included,including 9 males and 6 females,aged 48-72 years with an average age of(53.2 ± 6.5)years.The course of disease was(7.47 ± 3.45)years,ranging from 4 to 12 years;the preoperative H-Y grade was(3.1 ± 0.9),ranging from 2 to 4.5.All the 15 patients were reexamined with cranial CT after operation.The deviation between the electrode tip and the planned target on x-axis and y-axis was compared.The maximum deviation distance of x-axis was 1.09 mm,and the average deviation was(0.62 ± 0.23)mm;the maximum deviation distance of Y axis was 0.95 mm,and the average deviation distance was(0.53 ± 0.26)mm.15 patients had a total score of UPDRS III of off-drug states(49.8 ± 10.2),a total score of UPDRS III of on-drug states(31.6 ± 10.8)before operation,a total score of UPDRS III of sim on+drug off states(32.8 ± 12.5)and a total score of UPDRS III sim on+drug on states(22.4 ± 9.6)after operation.Before the operation,the standard medoba tablet was used for acute levodopa shock test.The effective time was(50.2 ± 7.4)min and the lasting effective time was(153.4 ± 42.6)min.At 12 months after operation and stim-on states,the effective time was shortened to(17.3 ± 4.7)min and extended to(190.2 ± 60.8)min after taking medpa.The equivalent total dose of levodopa was(820.8 ± 210.6)Mg on the day before operation and(440.6 ± 150.2)Mg on the day one year after operation.One year after operation,the total score of stim on+drug on state UPDRS III was significantly higher than that of preoperative state,and the difference was statistically significant.The score of UPDRS III in the state of stim on + drug off 1 year after operation can obtain the same results as the best pre-operative drug-on status.One year after operation,the effective time of levodopa was significantly shorter than that before operation,but the effective time was longer.The difference was statistically significant.One year after operation,the equivalent daily dose of levodopa was significantly lower than that before operation,and the difference was statistically significant.There was no significant difference in the cognitive and emotional function before and after operation.Conclusion: In this study,we found that the electrode can be accurately implanted with the help of frame stereotactic technology,and whether the electrode deviation can be accurately judged by the fusion of postoperative head CT and preoperative MRI plan images.Deep brain stimulation can improve the motor symptoms and quality of life of patients with moderate and severe Parkinson’s disease,and can significantly reduce the use of Parkinson’s disease drugs.After operation,the onset time of Parkinson’s disease drugs is significantly shortened and the duration is prolonged.There was no significant effect of operation on cognition,emotion,anxiety,depression and so on.There was uncertainty on the improvement of some non motor symptoms of Parkinson’s disease.The second part: The application of stereotactic technique in the model of Parkinson’s disease in mice: mi R-let-7a suppresses a-Synuclein-induced inflammation through targeting STAT3 in Parkinson’s diseaseBackground and Objective: The onset of Parkinson’s disease is a slow,long-term process,characterized by three major pathologies,including the progressive loss of dopaminergic neurons in the substantia nigra dense area,the formation of Lewy body(LB),and chronic neuroinflammation.The clinical manifestations of Parkinson’s disease include non-motor symptoms and motor symptoms.Non-motor symptoms appear early in the course of Parkinson’s disease,manifested by abnormal smell,pain,depression and anxiety symptoms,and may be accompanied by mental symptoms such as hallucinations.As the disease progresses and worsens,typical motor and systemic symptoms begin to appear,including resting tremor,bradykinesia,muscle stiffness,and postural balance disorders.Parkinson’s disease develops into an advanced stage.At present,it is still clinically focused on controlling symptoms and alleviating disease progression.There is a lack of treatment measures for the etiology and pathogenesis of Parkinson’s disease,which cannot protect dopaminergic neurons,reverse damaged dopaminergic neurons,and delay the progression of Parkinson’s disease.In the end,it causes Parkinson’s disease developing into a high disability rate at a later stage,which seriously affects the quality of life of patients and places a huge burden on society.Microglia-mediated neuroinflammation is an important risk factor for Parkinson’s disease progression and is closely related to degenerative lesions of dopaminergic neurons.The α-synapsin(α-Syn)is the main structural component of Lewy bodies,and its aggregation can activate the inflammation of microglia,leading to the occurrence and development of Parkinson’s disease.This study explored the application of stereotactic techniques in a mouse model of Parkinson’s disease,and conducted in vitro experiments on how mi R-let-7a participated in the pathological development of Parkinson’s disease and the molecular mechanism of inhibiting α-Syn-induced neuroinflammation,which would provide new directions for the prevention and treatment of Parkinson’s disease.Methods: The mice were randomly divided into control group and Parkinson’s model group and there were 12 mice in each group.The Parkinson’s model group was injected with AAV2-α-syn PBS into the right substantia nigra par compactat(SNpc)of mice by stereotactic technique.The control group was injected with AAV2-α-vector PBS into the right SNpc.Three weeks later,the mi R-let-7a levels,the signal transducer and activator of transcription 3(STAT3)levels in SNpc of two groups were compared by real-time quantitative PCR(q RT-PCR).To explore the role of mi R-let-7a in regulating inflammation of Parkinson’s disease,the mice were randomly divided into NC mimic group and mi R-let-7a mimic group.Each group included 12 mice transplanted by AAV2-α-vector PBS and 12 mice injected with AAV2-α-syn PBS.The right corpus striatum of the mice were injected with NC mimic PBS or mi R-let-7a mimic PBS respectively 24 hours after the disease models.Three weeks after the Parkinson’s model was built,q RT-PCR was used to test the mi R-let-7a level,TNF-α,IL-6,IL-1β and IL-12 levels in the SNpc of two groups.And the STAT3、INOS and Iba-1 level of 2 groups was detected by western blot technology.Before the Parkinson model were build and three weeks after modelling,the traction test and MWM test were used to evaluate the traction ability and spatial learning and memory ability of the two groups of mice.Results: Three weeks after Parkinson’s model were build with AAV2-α-Syn,the expression level of mi R-let-7a in the SNpc of model mice was significantly lower than that of the control group.Contrary to the results of reduced mi R-let-7a levels,the expression levels of STAT3 and its phosphorylated protein(p-STAT3)were significantly upregulated in the SNpc of Parkinson’s model mice.In the mi R-let-7a mimic group,the mi R-let-7a levels of the control and model mice were highly expressed.Furthermore,the STAT3 activation and the up-regulation i NOS and Iba-1 levels induced by α-Syn were suppressed by the overexpression of mi R-let-7a.In addition,the transcript levels of TNF-α,IL-6,IL-1β and IL-12 in mice SNpc in the mi R-let-7a mimic group were lower than those in the NC mimic group(P <0.01).And the dyskinesia and spatial memory deficit in the mi R-let-7a mimic group was significantly improved compared with the NC mimic group(P <0.01).Conclusion: The expression levels between mi R-let-7a and STAT3 in the α-Syn-induced mice Parkinson’s model is in the opposite pattern.The STAT3-dependent Parkinson disease inflammation reaction could be suppressed by mi R-let-7a.In addition,the dyskinesia and spatial memory deficits in Parkinson model mice could also be improved by mi R-let-7a mimic transplantation.